SP13. Early-Onset Metformin Regimen Inhibits Radiation-induced Fibrosis in Mice Model

Abstract

mouse anti-canine FGF7 (1:50 dilution). Primary tenocyte culture and treatment conditions: FGF7 and Periostin are two proteins that correlate to FGF7 and POSTN genes. They were chosen to test their functional effects on canine FDP tenocyte biology. Tenocytes were classified into 4 groups: non-treated control group, FGF7 group, periostin group, and FGF7 with periostin group. Tenocyte proliferation assay: The cells were incubated with completed medium (DMEM supplemented with 10% FBS, control) containing 0.5ug/ml FGF7 and/or 1ug/ml Periostin. Images were captured by the Incucyte® Live-Cell Analysis System. Caspase-3/7 mediated apoptosis assay: The Incucyte ® Caspase-3/7 Green Dyes were diluted to 1:1000 in full culture medium. Cells were then treated with 0.5ug/ml FGF7 and/or 1ug/ml Periostin in MEM medium containing Caspase-3/7 Green Dyes. The Caspase-3/7 green apoptosis assay reagent had the ability to show green fluorescent staining of nuclear DNA after being cleaved by Caspase-3/7. RESULTS: Differentially expressed (17826) genes were screened in the chordae tendineae compared to the flexor tendon, among which, 1172 genes with significantly increased expression (log2 fold change > 2) and 547 with decreased expression (log2 fold change < 2). The 20 top up and down-regulated genes are noted. Moreover, we found that mRNA levels of POSTN and FGF7 were highly expressed in chordae tendineae, while almost no expression was observed in flexor tendon (P<.05). Immunohistochemical analyses also confirmed this finding. The caspase-3/7 apoptosis assay showed promising effects on anti-apoptosis of the proteins. CONCLUSIONS:This preliminary discovery promotes a novel idea for tenogenesis using specific molecules identified from chordae tendineae in the future.